Chemoenzymatic synthesis of complex N-glycans of the parasite S.mansoni to examine the importance of epitope presentation on DC-SIGN recognition

Apoorva D Srivastava; Luca Unione; Mehman Bunyatov; Ivan A Gagarinov; Nicola G A Abrescia; Sandra Delgado; Ana Arda; Geert-Jan Boons

doi:10.1002/anie.202105647

Chemoenzymatic synthesis of complex N-glycans of the parasite S.mansoni to examine the importance of epitope presentation on DC-SIGN recognition

Apoorva D Srivastava
, Luca Unione
, Mehman Bunyatov
, Ivan A Gagarinov
, Nicola G A Abrescia
, Sandra Delgado
, Ana Arda
, Geert-Jan Boons

CIC BioGUNE

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

The importance of multivalency for N-glycan-protein interactions has primarily been studied by attachment of minimal epitopes to artificial multivalent scaffold and not in the context of multi-antennary glycans. N-glycans can be modified by bisecting GlcNAc, core xylosides and fucosides, and extended N-acetyl lactosamine moieties. The impact of such modifications on glycan recognition are also not well understood. We describe here a chemoenzymatic methodology that can provide N-glycans expressed by the parasitic worm S. mansoni having unique epitopes at each antenna and containing core xyloside. NMR, computational and electron microscopy were employed to investigate recognition of the glycans by the human lectin DC-SIGN. It revealed that core xyloside does not influence terminal epitope recognition. The multi-antennary glycans bound with higher affinity to DC-SIGN compared to mono-valent counterparts, which was attributed to proximity-induced effective concentration. The multi-antennary glycans cross-linked DC-SIGN into a dense network, which likely is relevant for antigen uptake and intracellular routing.

Original language	English
Pages (from-to)	19287-19296
Number of pages	10
Journal	Angewandte Chemie-International Edition
Volume	60
Issue number	35
Early online date	14 Jun 2021
DOIs	https://doi.org/10.1002/anie.202105647
Publication status	Published - 23 Aug 2021

Bibliographical note

Funding Information:
This research was supported by the Netherlands Organization for Scientific Research (NWO; TOP-PUNT grant 718.015.003 to G.J.B.), the Human Frontier Science Program Organization (HFSP; grant LT000747/2018-C to L.U.), Ram?n y Cajal (RYC) Contract (to A.A), the Agencia Estatal Investigacion of Spain (AEI; grants RTI2018-095700-B-I00 (to NGAA) and RTI2018-094751-B-C21) and the Severo Ochoa Excellence Accreditation (SEV-2016-0644). The staff of the Electron Microscopy Platform at the CIC bioGUNE and David Gil-Carton are acknowledged for valuable technical assistance. Dr. Sean Connell is acknowledged for the support in the 2D classification of DC-SGN ECD tetramers (EM). Dr. Prof. Jes?s Jim?nez-Barbero is acknowledged for insightful discussions, suggestions and comments.

Funding Information:
This research was supported by the Netherlands Organization for Scientific Research (NWO; TOP‐PUNT grant 718.015.003 to G.J.B.), the Human Frontier Science Program Organization (HFSP; grant LT000747/2018‐C to L.U.), Ramón Cajal (RYC) Contract (to A.A), the Agencia Estatal Investigacion of Spain (AEI; grants RTI2018‐095700‐B‐I00 (to NGAA) and RTI2018‐094751‐B‐C21) and the Severo Ochoa Excellence Accreditation (SEV‐2016‐0644). The staff of the Electron Microscopy Platform at the CIC bioGUNE and David Gil‐Carton are acknowledged for valuable technical assistance. Dr. Sean Connell is acknowledged for the support in the 2D classification of DC‐SGN ECD tetramers (EM). Dr. Prof. Jesús Jiménez‐Barbero is acknowledged for insightful discussions, suggestions and comments. y

Publisher Copyright:
© 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH

Keywords

NMR
chemoenzymatic synthesis
cryo-EM
glycan
glycosyl transferase

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Angew Chem Int Ed - 2021 - Srivastava - Chemoenzymatic Synthesis of Complex N‐Glycans of the Parasite S mansoni to ExamineFinal published version, 2.84 MBLicence: CC BY-NC

Cite this

Srivastava, A. D., Unione, L., Bunyatov, M., Gagarinov, I. A., Abrescia, N. G. A., Delgado, S., Arda, A., & Boons, G.-J. (2021). Chemoenzymatic synthesis of complex N-glycans of the parasite S.mansoni to examine the importance of epitope presentation on DC-SIGN recognition. Angewandte Chemie-International Edition, 60(35), 19287-19296. https://doi.org/10.1002/anie.202105647

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abstract = "The importance of multivalency for N-glycan-protein interactions has primarily been studied by attachment of minimal epitopes to artificial multivalent scaffold and not in the context of multi-antennary glycans. N-glycans can be modified by bisecting GlcNAc, core xylosides and fucosides, and extended N-acetyl lactosamine moieties. The impact of such modifications on glycan recognition are also not well understood. We describe here a chemoenzymatic methodology that can provide N-glycans expressed by the parasitic worm S. mansoni having unique epitopes at each antenna and containing core xyloside. NMR, computational and electron microscopy were employed to investigate recognition of the glycans by the human lectin DC-SIGN. It revealed that core xyloside does not influence terminal epitope recognition. The multi-antennary glycans bound with higher affinity to DC-SIGN compared to mono-valent counterparts, which was attributed to proximity-induced effective concentration. The multi-antennary glycans cross-linked DC-SIGN into a dense network, which likely is relevant for antigen uptake and intracellular routing.",

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note = "Funding Information: This research was supported by the Netherlands Organization for Scientific Research (NWO; TOP-PUNT grant 718.015.003 to G.J.B.), the Human Frontier Science Program Organization (HFSP; grant LT000747/2018-C to L.U.), Ram?n y Cajal (RYC) Contract (to A.A), the Agencia Estatal Investigacion of Spain (AEI; grants RTI2018-095700-B-I00 (to NGAA) and RTI2018-094751-B-C21) and the Severo Ochoa Excellence Accreditation (SEV-2016-0644). The staff of the Electron Microscopy Platform at the CIC bioGUNE and David Gil-Carton are acknowledged for valuable technical assistance. Dr. Sean Connell is acknowledged for the support in the 2D classification of DC-SGN ECD tetramers (EM). Dr. Prof. Jes?s Jim?nez-Barbero is acknowledged for insightful discussions, suggestions and comments. Funding Information: This research was supported by the Netherlands Organization for Scientific Research (NWO; TOP‐PUNT grant 718.015.003 to G.J.B.), the Human Frontier Science Program Organization (HFSP; grant LT000747/2018‐C to L.U.), Ram{\'o}n Cajal (RYC) Contract (to A.A), the Agencia Estatal Investigacion of Spain (AEI; grants RTI2018‐095700‐B‐I00 (to NGAA) and RTI2018‐094751‐B‐C21) and the Severo Ochoa Excellence Accreditation (SEV‐2016‐0644). The staff of the Electron Microscopy Platform at the CIC bioGUNE and David Gil‐Carton are acknowledged for valuable technical assistance. Dr. Sean Connell is acknowledged for the support in the 2D classification of DC‐SGN ECD tetramers (EM). Dr. Prof. Jes{\'u}s Jim{\'e}nez‐Barbero is acknowledged for insightful discussions, suggestions and comments. y Publisher Copyright: {\textcopyright} 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH",

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Srivastava, AD, Unione, L, Bunyatov, M, Gagarinov, IA, Abrescia, NGA, Delgado, S, Arda, A & Boons, G-J 2021, 'Chemoenzymatic synthesis of complex N-glycans of the parasite S.mansoni to examine the importance of epitope presentation on DC-SIGN recognition', Angewandte Chemie-International Edition, vol. 60, no. 35, pp. 19287-19296. https://doi.org/10.1002/anie.202105647

Chemoenzymatic synthesis of complex N-glycans of the parasite S.mansoni to examine the importance of epitope presentation on DC-SIGN recognition. / Srivastava, Apoorva D; Unione, Luca; Bunyatov, Mehman et al.
In: Angewandte Chemie-International Edition, Vol. 60, No. 35, 23.08.2021, p. 19287-19296.

Research output: Contribution to journal › Article › Academic › peer-review

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AU - Srivastava, Apoorva D

AU - Unione, Luca

AU - Bunyatov, Mehman

AU - Gagarinov, Ivan A

AU - Abrescia, Nicola G A

AU - Delgado, Sandra

AU - Arda, Ana

AU - Boons, Geert-Jan

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PY - 2021/8/23

Y1 - 2021/8/23

N2 - The importance of multivalency for N-glycan-protein interactions has primarily been studied by attachment of minimal epitopes to artificial multivalent scaffold and not in the context of multi-antennary glycans. N-glycans can be modified by bisecting GlcNAc, core xylosides and fucosides, and extended N-acetyl lactosamine moieties. The impact of such modifications on glycan recognition are also not well understood. We describe here a chemoenzymatic methodology that can provide N-glycans expressed by the parasitic worm S. mansoni having unique epitopes at each antenna and containing core xyloside. NMR, computational and electron microscopy were employed to investigate recognition of the glycans by the human lectin DC-SIGN. It revealed that core xyloside does not influence terminal epitope recognition. The multi-antennary glycans bound with higher affinity to DC-SIGN compared to mono-valent counterparts, which was attributed to proximity-induced effective concentration. The multi-antennary glycans cross-linked DC-SIGN into a dense network, which likely is relevant for antigen uptake and intracellular routing.

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KW - NMR

KW - chemoenzymatic synthesis

KW - cryo-EM

KW - glycan

KW - glycosyl transferase

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